Transformation of lacZ using different promoters in the commercially important red alga, Gracilaria gracilis

Huddy, Suzanne M.; Meyers, Ann E.; Coyne, Vernon E.

doi:10.5897/ajb11.2641

Cited by 6 publications

(1 citation statement)

References 35 publications

(29 reference statements)

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“…Thalli of the red alga Kappaphycus alvarezii was used in the first report of the transient transformation of seaweeds using particle bombardment (Kurtzman and Cheney, 1991). Transient transformation systems have been established in the red macroalgae Gracilaria chagii, G. gracilis, and K. alvarezii by particle bombardment (Gan et al 2003a;Wang et al, 2010a;Huddy et al, 2012); the life cycle of the three organisms corresponds to heteromorphic alternation of generations. In the brown macroalgae, Laminaria japonica and Undaria pinnatifida, transient transformation has been described in sporophytes by particle bombardment (Daozhan et al, 2002;Jiang et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

A Toolbox for Constructing a Stable Genetic Transformation Platform Allowing Foreign Fragment Integration in the Genome of Neopyropia yezoensis

et al. 2022

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Neopyropia yezoensis, previously known as Pyropia yezoensis, is regarded as a model macroalgae because of its economic value and typical intertidal seaweed species. However, the genetic transformation system to introduce foreign genes into its cells is inefficient and not stable. This study developed a more efficient transformation toolbox that allows the stable expression of foreign genes in N. yezoensis cells. We constructed an efficient transformation platform with the parameters of biolistic bombardment of 6 cm target distance, 600 μg gold particles/shot, 10 μg plasmid DNA/shot, 1,350 psi of helium, and 30 mmHg vacuum pressure. Thalli at 35 days of age were the most suitable transformation conditions, in which the highest transformation efficiency was generated. The endogenous promoter pPyACT1 could control gene expression efficiently compared to pPyUBC, pPyDPE2, and pPyEF1-a, especially the exogenous promoter d35S. Finally, the foreign genes PyGUS and PyHygR were stably expressed in different generations of transformants, including monospores, gametophytes, and filamentous sporophytes. Southern blotting analysis confirmed that PyGUS was integrated into the genome of N. yezoensis transformants. Establishing an efficient gene expression toolbox provides a strong foundation for functional genomics research and molecular genetic breeding on N. yezoensis.

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Section: Introductionmentioning

confidence: 99%

A Toolbox for Constructing a Stable Genetic Transformation Platform Allowing Foreign Fragment Integration in the Genome of Neopyropia yezoensis

et al. 2022

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Molecular mechanism underlying Pyropia haitanensis PhHsp22-mediated increase in the high-temperature tolerance of Chlamydomonas reinhardtii

et al. 2021

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Global warming is one of the key limiting factors affecting the cultivation of Pyropia haitanensis which is an economically important macroalgae species grown in southern China. However, the mechanism underlying the high-temperature tolerance of P. haitanensis remains largely unknown. In a previous study, we showed that the expression of the small heat shock protein 22 gene (Hsp22) is upregulated in P. haitanensis in response to high-temperature stress, but the associated regulatory mechanism was not fully elucidated. In this study, a transgenic Chlamydomonas reinhardtii expression system was used to functionally characterize P. haitanensis Hsp22. Our analyses indicated that the C-terminal of PhHsp22 is highly conserved and contains an A-crystal structure domain. A phylogenetic analysis revealed PhHsp22 is not closely related to small heat shock protein genes in other species. Additionally, PhHsp22 expression significantly increased at 3 and 6 h after initiating 33 °C treatment, which improved the survival rate of transgenic C. reinhardtii during the early stage of high-temperature treatment. The further transcriptome analysis revealed that PhHsp22 expression can promote pathways related to energy metabolism, metabolites metabolism, and protein homeostasis in transgenic C. reinhardtii cells exposed to high temperatures. Therefore, PhHsp22 may be crucial for the response of Pyropia species to high-temperature stress. Furthermore, this gene may be useful for breeding new high-temperature algal strains.

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Functional genomic and transformation resources for commercially important red macroalgae (Rhodophyta)

Khatri,

Patel,

Adams

et al. 2023

Algal Research

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Transformation of lacZ using different promoters in the commercially important red alga, Gracilaria gracilis

Cited by 6 publications

References 35 publications

A Toolbox for Constructing a Stable Genetic Transformation Platform Allowing Foreign Fragment Integration in the Genome of Neopyropia yezoensis

A Toolbox for Constructing a Stable Genetic Transformation Platform Allowing Foreign Fragment Integration in the Genome of Neopyropia yezoensis

Molecular mechanism underlying Pyropia haitanensis PhHsp22-mediated increase in the high-temperature tolerance of Chlamydomonas reinhardtii

Functional genomic and transformation resources for commercially important red macroalgae (Rhodophyta)

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